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RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...

Next-generation Sequencing

Next-generation Sequencing

The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
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Sanger Sequencing

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...

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Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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Related Experiment Video

Updated: May 19, 2026

Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins

Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins

Published on: August 9, 2019

Technical considerations for functional sequencing assays.

Weihua Zeng¹, Ali Mortazavi

¹Department of Developmental and Cell Biology, University of California, Irvine, California, USA.

Nature Immunology

|August 23, 2012

Summary

This summary is machine-generated.

Careful experimental design and result evaluation are crucial for next-generation sequencing (NGS) technologies. This discussion covers technical considerations, quality assessment criteria, and validation for high-throughput assays.

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Last Updated: May 19, 2026

Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins

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Next Generation Sequencing for the Detection of Actionable Mutations in Solid and Liquid Tumors

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Area of Science:

Genomics
Molecular Biology
Biotechnology

Background:

Next-generation sequencing (NGS) technologies are powerful tools in biological research.
Meeting the stringent requirements of various scientific fields necessitates rigorous experimental design and data evaluation.

Purpose of the Study:

To outline key technical considerations for designing and executing high-throughput sequencing experiments.
To provide criteria for assessing the quality of NGS results.
To highlight the importance of validation in ensuring reliable data.

Main Methods:

Discussion of experimental design principles for high-throughput assays.
Presentation of quality control metrics for sequencing data.
Review of validation strategies for NGS applications.

Main Results:

Identification of critical technical factors influencing NGS assay performance.
Establishment of benchmarks for evaluating sequencing data quality.
Emphasis on the necessity of robust validation protocols.

Conclusions:

Optimizing experimental design and data evaluation is essential for successful NGS implementation.
Adherence to quality assessment and validation criteria ensures the reliability and reproducibility of high-throughput sequencing results.